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Biomedical subjects

F Fish

Publications and source records attributed to F Fish.

At least 19 recordsLinked to original sources
PubMed1999

Congenital long-QT syndrome caused by a novel mutation in a conserved acidic domain of the cardiac Na+ channel.

BACKGROUND: Congenital long-QT syndrome (LQTS) is an inherited condition of abnormal cardiac excitability characterized clinically by an increased risk of ventricular tachyarrhythmias. One form, LQT3, is caused by mutations in the cardiac voltage-dependent sodium channel gene, SCN5A. Only 5 SCN5A mutations have been associated with LQTS, and more work is needed to improve correlations between SCN5A genotypes and associated clinical syndromes. METHODS AND RESULTS: We researched a 3-generation white family with autosomal dominant LQTS who exhibited a wide clinical spectrum from mild bradycardia to sudden death. Molecular genetic studies revealed a single nucleotide substitution in SCN5A exon 28 that caused the substitution of Glu1784 by Lys (E1784K). The mutation occurs in a highly conserved domain within the C-terminus of the cardiac sodium channel containing multiple, negatively charged amino acids. Two-electrode voltage-clamp recordings of a recombinant E1784K mutant channel expressed in Xenopus oocytes revealed a defect in fast inactivation characterized by a small, persistent current during long membrane depolarizations. Coexpression of the mutant with the human sodium channel beta1-subunit did not affect the persistent current, even though we did observe shifts in the voltage dependence of steady-state inactivation. Neutralizing multiple, negatively charged residues in the same region of the sodium channel C-terminus did not cause a more severe functional defect. CONCLUSIONS: We characterized the genetics and molecular pathophysiology of a novel SCN5A sodium channel mutation, E1784K. The functional defect exhibited by the mutant channel causes delayed myocardial repolarization, and our data on the effects of multiple charge neutralizations in this region of the C-terminus suggest that the molecular mechanism of channel dysfunction involves an allosteric rather than a direct effect on channel gating.

Adolescent↗
PubMed1997

Opercular jetting during fast-starts by flatfishes

When attacked by predators, flatfishes perform fast-starts that result in a rapid take-off from the ocean bottom on which they lie. High-speed video recordings of the blind side of flatfishes indicate that they expel a coherent jet of water from the blind-side opercular valve during take-off. Buccal pressure recordings in winter flounder (Pseudopleuronectes americanus) show that a buccal pressure pulse begins 0­20 ms before the beginning of the fast-start and has a range of mean magnitudes for three individuals of 1.6­10.7 kPa. We hypothesize that one function of the opercular jet in flatfishes may be to reduce the effects of Stefan adhesion. Stefan adhesion occurs as the fish lifts its head up rapidly from the ocean bottom, when water must flow into the space forming beneath the fish. Water viscosity opposes this rapid shear, and a suction pressure develops under the fish, making it more difficult for the fish to escape from the bottom. To estimate the magnitude of Stefan adhesion, we simulated fast-starts using a physical model in which a dead flounder was pulled upwards with an acceleration of 95 m s-2. Results from the physical model indicate that up to 35 % of the total force required to lift the head at 20 ms into the start can be attributed to Stefan adhesion. Despite this large adhesion force, previous work has shown that live flatfish do not show improved fast-start performance when Stefan adhesion has been eliminated by starting the fish from an open wire grid. Thus, live fishes are likely to be using behavioral mechanisms to reduce the adhesion force. Both the timing and location along the body of the opercular jet indicate that it is ideally suited to attenuate the effects of Stefan adhesion. Propping the body up on the median fins may also reduce adhesion by increasing the initial distance between the fish and the ocean floor.

Journal Article↗
PubMed1995

Hydroplaning by ducklings: overcoming limitations to swimming at the water surface

Rapid escape behavior by mallard (Anas platyrhynchos) ducklings is restricted to burst swimming at the water surface. Maximum speed may be limited because of the pattern of waves created as the duckling's body moves through the water (hull speed). Burst speeds for 9-day-old ducklings were compared with predicted hull speeds, based on the waterline length of ducklings either resting in water or actively swimming. Kinematic analysis of video tapes showed a mean maximum burst speed of 1.73 m s-1, which was four times greater than the predicted hull speed. At burst velocities, stroke frequency was 1.9 times higher than the stroke frequency measured during steady low-speed paddling. Transition to burst speeds from steady paddling occurred near predicted hull speed. The paddling motions of the webbed feet were used to generate both thrust and lift. By using lift to raise the body above the water surface, the influence of waves in restricting maximum swimming speed is negated. The duckling's body becomes a planing type of hull and skims on the water surface.

Journal Article↗
PubMed1995

MR magnetization transfer measurements in temporal lobe epilepsy: a preliminary study.

MR magnetization transfer ratio was measured in both hippocampi of three patients with temporal lobe epilepsy, and in two control subjects. The magnetization transfer ratio in each section was significantly lower on the affected side than on the contralateral side and in control subjects. Magnetization transfer ratio measurements are relatively fast and precise, this preliminary study shows that they may provide useful presurgical information.

Adult↗
PubMed1992

Analysis of the two-state behavior of the thermal unfolding serum retinol binding protein containing a single retinol ligand.

Through the use of CD and DSC, the thermal unfolding of holo serum retinol binding protein containing a single, tightly bound retinol ligand was studied at pH 7.4. The DSC endotherm of the holoprotein ([retinol]/[protein] = 1) was asymmetric about the transition temperature of 78 degrees C. Using changes in ellipticity at 230 nm, the thermal unfolding curve was also asymmetric about the inflection point centered near 78 degrees C. van't Hoff enthalpies were determined by three means and compared to the calorimetric enthalpy (delta Hcal) of 200 kcal/mol. A van't Hoff enthalpy of 190 kcal/mol was determined from the dependence of transition temperature on the concentration of the ligand-bound protein. This value agreed well with the van't Hoff enthalpies found from fits of the DSC (delta HvH = 184 kcal/mol) and spectroscopic (delta HvH = 181 kcal/mol) curves to a two-state thermodynamic model that included ligand dissociation (NR in equilibrium with U+R, where NR is the native holoprotein, U is the unfolded apoprotein, and R is retinol). Poor agreement was obtained with a two-state model that ignored ligand dissociation (N in equilibrium with U). Furthermore, the NR in equilibrium with U+R model accounted for the asymmetry in both CD and DSC transitions and yielded a much improved fit of the data over the N in equilibrium with U model. From these considerations and simulations on other equilibrium models, it is suggested that the NR in equilibrium with U+R model is the simplest model that describes the thermal unfolding of this ligand-bound protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Binding Sites↗
PubMed1988

Detection of genotoxic substances in cancer patients receiving antineoplastic drugs.

A short-term bacterial mutation test, the SOS Chromotest, has been used to detect the excretion in urine of genotoxic metabolites of antineoplastic drugs administered to cancer patients. In this test, the damage to the DNA of the test bacteria is expressed by the production of beta-galactosidase, which can be quantitatively assessed and is proportional to the concentration of the drug. Kinetic curves of excretion for adriamycin, bleomycin, dacarbazine, cis-platinum and vincristine and their mixtures have been constructed from standard curves relating the intensity of the beta-galactosidase response to the concentration of drugs dissolved in normal urine. Comparative data on extraction and concentration of the drugs from urine or serum by means of selective resin or silica columns are presented.

Adenocarcinoma↗
PubMed1987

Hydrophobic adherence and phase variation in Bordetella pertussis.

The hydrophobicity of Bordetella pertussis was assayed by measuring the ability of cells in suspension to adhere to a polystyrene surface. The quantity of adhered bacteria was measured by the binding of enzyme-conjugated anti B. pertussis antibodies. Hydrophobic adherence of non-pathogenic variant strains was about 20% of that exhibited by pathogenic strains. Hydrophobicity was a stable trait as it did not change with passaging or storage. Assays of a series of characterized stable variants suggested that the Filamentous Hemagglutinin (FHA) is the cell surface moiety responsible for hydrophobic adherence in B. pertussis.

Bacterial Adhesion↗
PubMed1987

Phase variation in Bordetella pertussis is accompanied by changes in DNA modification.

Pathogenic strains of Bordetella pertussis tend to undergo a phase variation process when propagated in vitro. The phase variants do not express part or all of the virulence factors of the pathogenic strain and are phenotypically stable. We have previously shown that variation involves a non-reversible, non-random process. In an attempt to characterize the molecular changes accompanying phase variation, chromosomal DNA, isolated from B. pertussis and its variants, was digested with a variety of restriction enzymes followed by agarose gel electrophoresis. While variant DNA was digested by all tested enzymes, pathogenic strain DNA was not digested by part of the enzymes, thus suggesting modification of the DNA at specific sites. DNA isolated from reversible, growth medium induced variants, demonstrated sensitivity to digestion identical to that of spontaneous, stable variants. Analysis of the restriction sequences of all the enzymes, which did not digest DNA from pathogenic strains, failed to reveal any common sequence known to be affected by methylation. HPLC and nearest-neighbor analysis showed a 2-fold increase in the level of DNA methylation in the pathogenic strain. It was concluded that (a) the chromosomal DNA in virulent strains of B. pertussis is protected against enzymatic digestion by an as yet unknown modification and (b) variation in B. pertussis may be caused by changes in the modification of the DNA rather than by mutation.

Base Sequence↗
PubMed1987

The detection of neurotoxicant activity by a bacterial toxicity assay.

We present and evaluate a simple, rapid (2 hrs), colorimetric assay for the detection of toxicants, including neurotoxicants. The assay is based on the ability of toxicants to inhibit the de novo synthesis of an inducible enzyme, beta-galactosidase, by a rough mutant of E. coli, which is highly sensitive to a wide spectrum of toxic substances. The test is performed under stress conditions for the bacterium, since under such conditions better sensitivity for low concentrations of analytes is obtained. In the assay, the sample is mixed with the stressed bacteria and a cocktail containing the specific inducer for the chromogenic enzyme and factors essential to the recovery of the bacteria from their stressed condition. The ability of cells to synthesize B-galactosidase under these conditions depends on their ability to recover from the stress. Toxic materials interfere and/or inhibit the recovery process and with it the synthesis of beta-galactosidase.

Animal Testing Alternatives↗
PubMed1985

Studies on phase variation in Bordetella pertussis.

Pathogenic strains of Bordetella pertussis undergo spontaneous phase variation and become non-pathogenic upon culturing in vitro. The spontaneous process was studied in pathogenic B. pertussis strains Tohama, 165 and 18323 by isolating spontaneous variants, selected for their ability to grow on synthetic and semi-synthetic solid media. In strains Tohama and 165, the frequency of variants able to grow on synthetic and semi-synthetic media was between 10(-6) and 10(-7). About 250 variant strains were screened for the presence of virulence-associated traits, such as production of hemolysin, pertussis toxin and filamentous hemagglutinin (FHA). Only four different combinations of the traits were found: 7-11% of the variants displayed all traits, 17% of the variants carried pertussis toxin and FHA, 5-11% carried FHA only and 66% were devoid of all virulence traits. The strains which had at least one virulence trait also demonstrated some adenylate cyclase activity. The disappearance of hemolysin was related quantitatively to the other traits. These results suggest that phase variation in B. pertussis is a non random process, involving ordered disappearance of virulence factors in the following order: hemolysin, pertussis toxin and FHA. Since all the variant strains were phenotypically stable upon further passaging in vitro, they represent the stable, final outcome of the variation process which may have occurred in "Phase I" colonies. In contrast, 300 variants of B. pertussis 18323, which were able to grow on selective solid media, carried all the virulence traits. This is in accordance with the strain's unique intracerebral virulence.

Adaptation, Physiological↗
PubMed1984

Spontaneous phase variation in Bordetella pertussis is a multistep non-random process.

Pathogenic strains of Bordetella pertussis undergo spontaneous phase variation and become non-pathogenic upon culturing in vitro. Spontaneous variants of the Tohama and #165 pathogenic strains of B. pertussis were selected by their ability to grow on synthetic and semi-synthetic solid media. The frequency of these variants was between 10(-6) and 10(-7). About 250 variant strains were screened for the presence of virulence-associated traits, such as production of hemolysin, pertussis toxin and filamentous hemagglutinin (FHA). Only four different combinations of the traits were found: 7-11% of the variants displayed all traits, 17% of the variants carried the toxin and FHA, 5-11% carried FHA only and 66% were devoid of all virulence traits. The strains which had at least one virulence trait also demonstrated some adenylate cyclase activity. The disappearance of hemolysin quantitatively affected the other traits. These results suggest that phase variation in B. pertussis is a non-random process, involving multistep disappearance of virulence factors in the following order: hemolysin, pertussis toxin and FHA. In contrast, all 300 variants of strain #18323 of B. pertussis, which were able to grow on the selective solid media, carried all the virulence traits. This is in accordance with the strain's unique intracerebral growth capability.

Adenylate Cyclase Toxin↗
PubMed1984

Proliferative response of immune mouse T-lymphocytes to the lymphocytosis-promoting factor of Bordetella pertussis.

Immunization of mice with a whole-cell pertussis vaccine or with the purified, detoxified lymphocytosis-promoting factor (LPF) of Bordetella pertussis resulted in an increased in vitro proliferative response to LPF in immune lymph node cells. The proliferative response was detected above the nonspecific mitogenic activity of LPF. That the proliferative response of the immune lymph node cells was a demonstration of a specific cell-mediated immunity to LPF was supported by the following: (i) the specificity of the response to the immunizing antigen; (ii) the ability of chemically modified, nonmitogenic LPF to induce proliferation in immune lymph node cells; and (iii) a dependence on T-cells for the demonstration of the proliferative response of immune cells to LPF. Immunization of mice with protective doses of detoxified LPF resulted in serum antibody and cell-mediated responses to LPF. Immunization of mice with protective doses of whole-cell pertussis vaccine resulted in a cell-mediated response but not a detectable antibody response to LPF. The LPF of B. pertussis is believed to play an important role in pathogenesis and immunity in pertussis, and the demonstration of a cell-mediated immune response to LPF suggests a possible role for cell-mediated immunity to LPF in protection from pertussis disease.

Animals↗
PubMed1983

Pertussis toxin. Affinity purification of a new ADP-ribosyltransferase.

Pertussis toxin, the major toxin produced by Bordetella pertussis, catalyzes the ADP-ribosylation of a specific membrane polypeptide which appears to be involved in regulation of the catalytic subunit of adenylate cyclase. In the current study, a rapid purification procedure has been developed for the preparation of pertussis toxin in high yields. Through the sequential use of the affinity matrices Affi-Gel blue and fetuin-Sepharose 4B, milligram quantities of apparently homogeneous toxin can be prepared from the culture supernatants of B. pertussis strain 165. Structural, amino acid, and immunologic analyses indicate that toxin prepared from strain 165 is indistinguishable from toxin prepared from other strains. Activation of the ADP-ribosyltransferase activity requires treatment of the toxin with a thiol reducing agent. This activation appears to be associated with the reduction of intrachain disulfide bonds present in the catalytic subunit. Activated toxin preparations catalyzed ADP-ribosylation of a protein (Mr = 40,000) present in cell membrane preparations obtained from human red blood cells and platelets, rat adipocytes, and cyc- S49 cells which are deficient in the adenylate cyclase regulatory component which is the substrate for cholera toxin.

Adenylate Cyclase Toxin↗
PubMed1982

Hidden anti-double stranded DNA antibodies in autoimmune mice.

When MRL/l mouse spleen cell culture supernatants were incubated with normal mouse spleen cells, a two-50-fold increase in anti-dsDNA activity was noted. A smaller increase in anti-ssDNA and no change in anti-TNP antibody activity were observed. This 'hidden' antibody in the MRL/l supernatants could not be revealed by DNAse digestion and could not be absorbed by a DNA cellulose column. Hidden antibody was removed from supernatants by sepharose-anti Ig. After DNAase digestion of the MRL/l supernatants, hidden anti-dsDNA could not be revealed by incubation with spleen cells. All the hidden activity was excluded by gel filtration on Sephacryl S-300 (mol. wt greater than 300,000) but was banded in the low density protein area of caesium chloride equilibrium density gradients. It was also noted that MRL/l mouse sera had hidden anti-dsDNA antibodies. Hidden antibodies were present in both the IgG and IgM classes. The revealed antibodies demonstrated impaired ability to bind Fc specific anti-Ig reagents suggesting that they were partially degraded during the incubation with mouse spleen cells. The hidden anti-dsDNA thus appears to represent a DNA-anti-dsDNA complex, perhaps of very high affinity. It may explain why anti-dsDNA but not anti-ssDNA antibodies are of pathological importance in SLE.

Animals↗
PubMed1982

The in vitro and in vivo induction of anti-double-stranded DNA antibodies in normal and autoimmune mice.

To determine whether the existence of anti-dsDNA producing lymphocyte clones is limited to autoimmune strains of mice, spleen cells derived from autoimmune mice (NZB, NZB X NZW F1, MRL) and from normal strains (BALB/c, DBA/2, C57BL/6, C3H/eb) were cultured with E. coli lipopolysaccharide (LPS). DNase-treated supernatants from these cultures were assayed for anti-dsDNA antibodies by employing a sensitive solid-phase radioimmunoassay with poly (dA-dT) as the antigen. All tested spleen cells secreted a small yet significant amount of anti-dsDNA upon stimulation with LPS. There was no difference in the amount or in the heavy chain type of anti-dsDNA secreted by cells from normal and autoimmune strain cells. Evidence of clonal expansion in unstimulated cells was observed only in cultures prepared from older autoimmune animals. Removal of T cells from the spleen cell preparations had no marked effect on the spontaneous or stimulated antibody secretion. Anti-dsDNA antibodies could also be induced in vivo by i.p. injection of LPS into young normal animals. Splenocytes from all tested strains spontaneously secreted anti-ssDNA and anti-TNP antibodies in culture, and these were present at relatively high levels in the serum of unstimulated animals. Stimulation with LPS increased secretion of anti-ssDNA and anti-TNP in all strains in vitro and in five of seven strains in vivo as well. It can be concluded that a) the existence of anti-dsDNA-producing clones is not limited to autoimmune strains, and b) these clones are expanded in old but not in young autoimmune mice. They are not expanded in normal mice at any age.

Aging↗
PubMed1981

A sensitive solid phase microradioimmunoassay for anti-double stranded DNA antibodies.

A sensitive solid phase microradioimmunoassay has been developed for measurement of antidouble stranded DNA (dsDNA) antibodies. In this procedure, advantage has been taken of the capacity of poly-L-lysine (PLL) to facilitate the binding of pure dsDNA to plastic surfaces. In the absence of PLL, binding did not occur. Diluted sera were incubated in PLL-treated dsDNA-coated microtitration trays and anti-dsDNA Ig measured using affinity purified 125I-anti-Ig of high specific activity. The synthetic DNA, poly dA-dT, was used as a model for dsDNA. In initial experiments, specific anti-DNA binding could not be demonstrated because of high background binding of patient Ig to PLL-treated surfaces. This was reduced by diluting test sera and anti-Ig in buffer containing 2% BGG and 1% BSA. Specificity of the assay for DNA was demonstrated by absorbing the anti-DNA activity on DNA-coated plastic. The binding of systemic lupus erythematosus (SLE) patient serum Ig to poly dA-dT coated trays did not diminish after digestion with nuclease S1, suggesting that the synthetic polymer is an appropriate model for dsDNA. Patient and normal sera were screened for anti-dsDNA activity using poly dA-dT as antigen. None of the 38 normal sera, 23 of 35 active SLE sera, 1 of 25 treated SLE, 4 of 35 rheumatoid arthritis, 3 of 35 scleroderma, and 1 of 13 polymyositis sera demonstrated positive anti-dsDNA activity. The anti-dsDNA values obtained in the radioimmunoassay correlated significantly with those obtained in the Crithidia luciliae assay.

Antibodies↗